Sounds too easy, but I'm having problems with changing the ampicillin
resistance of a pUC119 based plasmid with any other resistance. I am
cutting with BspH1, which flanks lactamase, and trying to clone in a PCR
fragment from Alexeyev's cassette antibiotic resistances (chloramphenicol,
gentamycin or kanamycin). I get resistant clones, but they are all
rearranged. Is there something special about the bit between the two BspH1
sites? Any suggestions? Am I doing something silly?
I have been thinking of amplifying the plasmid, excluding amp, and just
adding sites to the end of the PCR product. Any suggestions about which
bits I should keep and which I should not?
thanks for your help
andrew bradbury
Andrew Bradbury
SISSA c/o ICGEB
Area di Ricerca
Padriciano 99
Trieste 34012
Italy
Tel +39 40 398995
Fax +39 40 398991